山羊卵母细胞体外培养体系及冻存后体外培养的研究

摘要

With the rapid development of the mammalian embryo engineering technology and commercialization of embryo transplantation technique, the demand for oocyte increases dramatically. The oocyte cryopreservation and in vitro culture system are very important steps in the process.In this study, the oocytes from Yunling goat were used to study its in vitro culture system.The results showed that the mauturation rates of the control (OM + 10 % FBS) and the group I with 1 % ITS (OM + 10 % FBS +1 % ITS) had no significant difference (71.6 % vs 76.2 %),while the cleavage rates of parthenogenetic activation in two groups were significantly different (60.5 % vs 71.5 % ).With optimal oocyte maturation system to cultivate thawing COCs in GV stage,the mature rate was 31.5 %.Cultivated the GV and Mil oocytes that were thawed and parthenogenetic activated in embryo culture medium with ITS and EGF.the rates of cleavage, were 35.6 % and 58.4 % respectively, had significant difference (P<0.01).%随着哺乳动物胚胎工程技术的深入研究和商业化胚胎移植技术的快速发展,对卵母细胞的需求量急剧增加.而冷冻的卵母细胞可为胚胎工程技术研究提供方便、丰富的材料来源.试验以屠宰场云岭黑山羊卵巢卵母细胞为实验材料,研究冷冻后的山羊卵母细胞体外培养体系.结果表明,采用传统成熟培养液(OM+10％FBS)与在其中添加1％ ITS(OM+ 10 ％FBS+1％ ITS)对卵母细胞培养成熟率无显著差异(71.6％vs 76.2％),而孤雌激活的卵裂率差异显著(60.5％vs 71.5％).用优化的卵母细胞成熟体系培养解冻后的GV期COCs,成熟率31.5％；用添加有ITS和EGF的胚胎培养液培养解冻后GV和MⅡ期孤雌激活的卵母细胞,其卵裂率分别为35.6％、58.4％,差异极显著(P＜0.01).